The present invention relates to enzymes, and more particularly, to an enzymatic composition and system for delivering same whereby long term stability and efficacy of enzymes contained therein may be maintained.
Enzymes are proteins which catalyze specific reactions. The number of different types of reactions that enzymes catalyze is enormous. The extraordinary range and specificity of reactions catalyzed by enzymes has led to their uses in almost all areas of technology, such as agriculture, manufacture of raw materials for the clothing, pharmaceutical and cosmetics industries, for chemical synthesis, etc. Each enzyme shows specificity for a substrate that ranges from fairly general to absolutely specific. For example, trypsin is a generally non-specific enzyme which hydrolyzes the peptide bond between an arginine and another amino acid in almost any protein. An example of a highly specific enzyme is Eco RI, which will hydrolyze the phosphodiester bond between guanidine and adenosine residues in DNA only if they are present in the specific sequence GAATTC. The enzyme (Eco RI) will not work in any other sequence, nor will it work if any of the bases of the sequence have been modified with a methylene group.
Proteolytic enzymes, or proteases, are a class of enzymes which will generally hydrolyze the peptide bond in proteins and thus convert proteins into polypeptides. A large number of proteolytic enzymes is known, each with a different amino acid specificity, e.g., trypsin, papain, subtilisin, interleukin-converting enzyme, collegenase, etc. Many of these proteins have been put to use in various beneficial ways, such as in cosmetic and medicinal formulations, pharmaceuticals, food products, etc. Difficulties in using proteolytic enzymes in such products arise, however, because a good many proteins (e.g., other enzymes) within such cosmetic and medicinal formulations may be affected by the enzyme (e.g., hydrolized by the protease action). Again, all enzymes are proteins the peptide bonds of which are susceptible to hydrolysis by proteolytic enzymes. Accordingly, chemists and/or formulation personnel preparing simple admixtures containing proteolytic enzymes and other active proteins are confronted with the problems associated with hydrolysis of the proteins, and the task of assuring long-term efficacy of the formulations.
The following emphasizes the problem which the present invention addresses. If one wanted a composition of matter which contained both trypsin and scrum albumin, a two container system or separation system would be required to protect the albumin from hydrolysis by the trypsin in the composition. Conventional two-container or microencapsulation systems, however, are difficult to accurately and efficaciously use. That is, they merely reduce or slow the proteolytic activity, i.e., the hydrolytic action of the protease on the non-proteolytic enzyme. Stability, therefore, of compositions of matter containing a protease is possible with conventional encapsulation technology only for the short term.
For example, U.S. Pat. No. 4,668,630 to Louderbach discloses a composition containing at least one enzyme. Substantially all of the one enzyme is combined with a stabilizing amount of reversible inhibitor. The inhibitor forms an inhibitor-enzyme complex thereby stabilizing the enzyme. While such methods reduce proteolysis and result in improved protein or enzyme stability, the efficacy of the proteolytic enzyme is compromised.
Further, U.S. Pat. No. 5,281,356 discloses another example of a two-container encapsulation system provided to separate or minimize interaction between non-compatible enzymes. The system embodies a composition comprising a non-proteolytic enzyme, such as lipase or cellulase, and capsules containing a proteolytic enzyme and a composite emulsion polymer. Were the proteolytic enzyme not encapsulated, its action would degrade the other constituent proteins or non-proteolytic enzymes. The emulsion polymer comprises a hydrophilic portion attached to hydrophobic core particles as a network to entrap the enzymes between the hydrophobic particles and preferably crosslinked water soluble polymer to act like a gel or sieve. The proteolytic enzyme diffusion through the same is thereby slowed, which in turn slows the rate of degradation of the non-proteolytic enzyme thereby. Such a composition, however, does not maintain efficacious separation.